6-chloro-1-dehydro steroid compounds



United States Patent 0 3,130,211 Patented Apr. 21, 1964 3 130 2112,838,532, and a division of application Serial No. 634,- 6 CHL0R0 1])EHYDR0 STEROID 828, filed January 18, 1957, now Patent 2,841,600.

CONIPOUNDS It has been discovered that 6-halo-1-dehydrohydro- Georgespew Kalamazoo Township, Kalamazoo cortisone and the 21-esters thereofpossess valuable anti c t Mj h i to The Upjnlm ,K 1 15 rheumatoidarthritic, anti-inflammatory and glucocortimazo0,lVIich.,aco1-p0rationof Michigan coid activity. The compounds are useful in the treat- NoDrawing. Original application Jan. 18, 1957, Ser. ment of inflammatoryconditions of the skin, eyes and 2? 110W Patel? dated y ears of humansand valuable domestic animals, contact Dlvlded and 11115 aPPllcatmn 19,1953 dermatitis and other allergenic reactions. The comg 260 397 20pounds can be administered in conventional dosage forms lmms' such aspills, tablets and capsules for oral use or in con- This inventionrelates to 6-halo-l-dehydrohydrocortventional liquid forms as are usedwith natural and isone and 21-esters thereof, to6-halo-1-dehydrocortisone synthetic cortical steroid hormones forinjection use. For and 21-esters thereof, to novel intermediates in thepro- 25 topical use they can be administered in the form of ointductionthereof and to processes for the production of ments, creams, lotionsand the like with or without cothe novel compounds and the novelintermediates. acting antibiotics, germicides and the like.

This application is a continuation-in-part of applica- The compounds ofthis invention can be prepared in tion Serial No. 519,632, filed July 1,1955, now Patent accordance with the following scheme of reactions:

CH3 CH3 (3003' 000R CH (1H II I R i R-fi CH CH ?II ?II CH0 GHO (CHalrO I(onaro II CH3 CH3 $003. $OOR' E R R? R CH3 CH3 RI! (EH-O I Qm-O E III EI Ha 5 X 5 X on on lLiAlHa CH3 CH3 $H2OH (I-JHZOAC l I HO HO OH OH R!IRI! (;3HO CHO (OHz)n0 I (arm-o i I V 5 X 3 X on on CH3 CH3 tllH OAcCHrOAc on :0 I "-013: HO- fii CH CH3 I I CH0 CHO i (emu-o I V (Cami-0 VIX i x s OH on CH5 CH3 $H OR OH OAe 0:0 0:0 L .OH OH H0 110?: 1

OH CH3 VIII 5 VII CH1 CH3 ornon omoR'" 0:0 0:0 L OH L--OH (\i fii /\CH3CH3 l I U o- I IX I X wherein R is fi-hydroxy or keto, R and R" arehydrogen or lower-alkyl, n is a whole number from one to two, X is ahalogen having an atomic weight from nineteen to eighty, inclusive,i.e., fluorine, chlorine or bromine, Ac is the acyl radical of ahydrocarbon carboxylic acid containing from one to twelve carbon atoms,inclusive, and R" is hydrogen or Ac as defined above. The termlower-alkyl, when used herein, refers to an alkyl radical of from one toeight carbon atoms, inclusive, e.g., methyl, ethyl, propyl, butyl, amyl,hexyl, heptyl, ethylhexyl, and octyl.

Oxidation of the product, 6-halo-1-dehydrohydrocortisone 21 acetate orother 21 esters, e.g., with chromic acid in acetic acid, is productiveof the 21- esters of 6-halo-l-dehydrocortisone, which compounds alsopossess glucocorticoid, anti-inflammatory and anti.- rhemutaoidarthritic activity.

The above reactions as described hereinbelow are embodiments of stepswhich will be exemplified in detail in the ensuing specification. Itwill be understood by those skilled in the art, nevertheless, that thespecific order of steps may be inverted or transposed or otherwisevaried to suit the purposes of economics, convenience, or the like.

The preferred compounds containing the 17(20)- double bond have the cisconfiguration, because the cis isomers can ultimately be converted inhigher yields in the oxidative hydroxylation step than is ordinarilypossible with the trans isomer. It should be understood, however, thatthe trans isomer or mixtures of the cis and trans isomers can be usedwith satisfactory results.

In carrying out the expoxidation step shown above, a S-ketal of 3-keto-11-oxygenated-4, 17 (20) -pregnadiene- Zl-carbonyloxy steroid representedby Formula I, which can be prepared as disclosed in US. Patent2,707,184, preferably the 3-ethylene glycol ketal of methyl3,11-diketo-4, 17 (20) [cis]-pregnadien-2l-oate, is epoxidized with aperacid, e.g., peracetic or perbenzoic, or other known epoxidizingagents, to produce the corresponding 5 :6-oxide (H). A mixture of boththe 04- and fi-oxides is produced in this epoxidation reaction, and themixture can be separated by chromatographic or crystallizationtechniques known in the art.

In the oxide opening step, a 3-ketalized3-keto-5a,6aoxido-11-oxygenated-17 (20) -pregnene 21 carbonyloxy steroid(II), is reacted with hydrogen fluoride, hydrogen chloride, or hydrogenbromide, preferably hydrogen fluoride, to open the oxide ring andproduce the corresponding 3-ketalized3-keto-5-hydroxy-6-halo-ll-oxygenatedl7(20)-pregnene-21-carbonyloxysteroid (Hi). This epoxide opening step is ordinarily carried out attemperatures between about minus forty and plus fifty degreesCentigrade, the preferred limits being between about zero and 25 degreescentigrade. It can be performed under anhydrous conditions in thepresence or absence of a catalyst, e.g.,-boron trifluoride; or underaqueous conditions in the presence or absence of a catalyst such as ahypohalous acid. Reaction conditions, for example, those disclosed bySchmidlin et al., Helv. Chim. Acta, 36, 1241 (1953); Gallagher, J. Biol,Chem., 162, 495 (1946); Cornforth et al., J. Chem. Soc., 1954, 907 andFried et al., J. Am. Chem. Soc., 75, 2273 (1953), are usually employed.

If anhydrous conditions are diflicult or inconvenient to maintain, theoxide opening reaction can be performed under aqueous conditions inwhich case the ketal will be hydrolyzed at the same time. Thethus-produced 3-keto group of Ila can then be Ieketalized in the mannerdescribed hereinabove to produce the corresponding ketal (HI).

In the reduction step of the present invention, a 3-ketalized 3keto--hydroxy-6-halo-1l-oxygenated 17(20)- pregene-Zl-carbonyloxysteroid (H1), preferably the 3- ethylene glycol ketal of lower-alkyl,preferably methyl 3,11 diketo 5-hydroxy-6-fluoro l7(20)-[cis]-pregnen-21-oate, is reduced with lithium aluminum hydride or other chemicalcarboxyl reducing agent in an organic solvent, e.g., ether, dioxane,tetrahydrofuran, benzene, to produce the corresponding 3-ketalized 5,11/3,2l-trihydroxy-6-halo-17(20)-pregnen-3-one. At completion of thisreaction, the reaction mixture is preferably mixed with water or, anacid, an ester or carbonyl agent followed by Water, to decompose anyexcess lithium aluminum hydride and organo-metal complexes. The usualreaction conditions for a lithium aluminum hydride reduction areemployed, except that a reaction temperature at room temperature orbelow is preferred, to ensure that reaction with the 6-halogen does notoccur, and acid, though operative and satifsactory under carefullycontrolled conditions, is preferably not employed in the decompositionstep, to avoid undue hydrolysis of the ketal group.

The esterification step, i.e., to produce Compound V, involves theconversion of a 21-hydroxy group of 5,113, 21trihydroxy-6-halo-17(20)-pregnen-3-one 3-alkylene ketal (IV) to a21-acyloxy group. This reaction can be performed under theesterification conditions known in the art, e.g., by the reaction of IVwith the selected acid halide or acid chloride or acid bromide or theanhydride of a hydrocarbon carboxylic acid, or by reaction with theselected acid, in the presence of an esterification catalyst or with anester under ester exchange reaction conditions. Reaction conditionswhich are apt to affect the labile 11B- hydroxy group or 6-halo groupshould be avoided. Compounds thus-produced include the compoundsrepresented by Formula V wherein the -17(20)-configuration is cis, the6-halogen is fluorine and the Ac is the acyl radical of a hydrocarboncarboxylic acid containing from one to twelve carbon atoms, inclusive,e.g., formic, propionic, butyric, isobutyric, valeric, isovaleric,trimethylacetic, 2- methylbutyric, 3-ethylbutyric, hexanoic,diethylacetic, triethylacetic, heptanoic, octanoic, a-ethylisovaleric, acyclic acid, e.g., cyclopropylideneacetic, cyclopentylformic,cyclopentylacetic, fl-cyclohexylpropionic, cyclohexylformic,cyclohexylacetic, an aryl or alkaryl acid, e.g., benzoic, 2-, 3- or4-methylbenzoic, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5- dimethylbenzoic,ethylbenzoic, 2,4,6-trimethylbenzoic, 2,4,6-triethylbenzoic,a-naphthoic, 3-methyl-a-naphthoic, an aralykyl acid, e.g., phenylacetic,phenylpropionic, diphenylacetic, triphenylacetic, etc.

The oxidative hydroxylation of V to V1 is carried out by reaction with acatalytic amount of osmium tetroxide and an oxidizing agent such ashydrogen peroxide, peracids, alkyl peroxides, amine oxide peroxides, andthe like. A preferred procedure involves the use of two to three molarequivalents of one of the oxidizing agents and less than 0.05 molarequivalent of osmium tetroxide, calculated on the basis of V, usingtertiary butyl alcohol as the reaction medium and carrying out thereaction at about room temperature.

The hydrolysis step (VI- VII) involves the hydrolysis of the ketal groupof the 3-ketalized 5,l1fi,17a-trihydroxy-6-halo-2l-acyloxypregnane-3,20-dione (VI), preferably the ethyleneglycol ketal thereof, employing aqueous acid to produce thecorresponding diketo compound VII. The hydrolysis is convenientlyperformed under relatively mild conditions, e.g., at room temperaturewith acetic acid or weak sulfuric acid.

The dehydration reaction is carried out using hydrogen chloride orGirards Reagent T. Acetic anhydride, ptoluene-sulfonic acid and aceticacid, or thionyl chloride and pyridine are also suitable as dehydratingagents. Care should be taken in this step to avoid various reactionconditions that would affect the l lfl-hydroxy group.

The l-dehydrogenation of VIII to 1X (where R is a 5- hydroxy group) canbe carried out microbiologically, i.e., fermentative l-dehydrogenationusing Septomyxa or Fusarium organisms. If loss of the 2l-ester takesplace during the fermentation, reacylation can be done by known21-acylation methods. Alternatively, the l-dehydrogenation can becarried out chemically in known manner by using selenium dioxide.

Alternatively, Compound VIH in the form of the 21- ester can be oxidizedto the corresponding ll-keto compound, i.e.,17a-hydroxy-6-halo-2l-acyloxy-3,l1,20-trione (Compound X), in accordancewith the process for the oxidation of the 21-esters of Kendalls CompoundF as disclosed in US. Patent No. 2,751,402. Compound X can be subjectedto l-dehydrogenation in the same manner as Compound VHI thus producingl7oc-hYdr0XY-6- halo-2l-acyloxy 1,4 pregnadiene 3,11, 20 trione (lX)where R is a keto group.

Either of Compounds VIII, IX or X can be transformed to the free21-alcohol by hydrolysis in accordance with known methods forhydrolyzing Compound F 21-esters to the free Compound F alcohol. Apreferred procedure is to employ at least a molar equivalent of analkali-metal bicarbonate in a substantially oxygen-free solution of amixture of a lower alkanol and water. The hydrolysis reaction is carriedout at a temperature between ten and thirty degrees centrigrade whileprotecting the mixture from atmospheric oxygen. After the hydrolysis iscomplete, the reaction mixture is neutralized with an acid, e.g., aceticacid, and the hydrolyzed product recovered from the reaction mixture byevaporation and crystallization, extraction with methylene chloride, orthe like.

As previously mentioned, the order of the various steps outlined abovecan be varied considerably. Suitable variations will occur to thoseskilled in the art, and the necessary adjustments can be readilyappreciated and carried out by skilled chemists. For example, CompoundIV can be hydrolyzed to remove the 3-ketal, and this step followed inturn by 2l-acylation, S-dehydration, oxidative hydroxylation andl-dehydrogenation under the above-described conditions.

The compounds represented by Formula IX and those represented byFormulas VIH and X, in addition to possessing the valuableanti-rheumatoid arthritic, anti-inflammatory and glucocorticoid activityheretofore mentioned, are additionally useful as intermediates for thepreparation of the 9a-halo analogues thereof, namely 9a,6dihalo17a,21-dihydroxy-4-pregnene-3,11,20-trione, 9a,6-dihalo-11,8,17a,21-trihydroxy 4 pregnene-3,20-dione including the corresponding21-esters thereof; and 9a,6-dihalo-17u,

2l-dihydroxy-1,4-pregnadiene-3,11,20-trione, and 9a,6-di.-halo-llfi,l7a,21-trihydroxy 1,4 pregnadiene-3,20-dione including thecorresponding 21-ester thereof. These 'di halo compounds also possessthe valuable anti-rheumatoid arthritic, anti-inflammatory andglucocorticoid activity of theparent '6-halo compounds, usually inhigher degree. They possess a combination of high anti-inflammatory and.glucocorticoid properties and at the same time possess lowmineralocorticoid properties.

In the preparation of these compounds, for example, 6fluoro-11'B,17u-dihydroxy 21 acetoxy-1,4-pregnadiene-3,20-d ione of theclass of Formula IX can be dehydrated with N br'omoacetamide andanhydrous sulfur dioxide until a negative acidified potassiumiodide-starch test of the reaction mixture is obtained. Dilution withcold water, e.g., ice water, results in the precipitation of'6-fluoro-17a-hydroxy-21-acetoxy 1,4,9(11) -pregnatriene-3,20-dionewhich can be purified with recrystallization from acetone. Thecrystalline product can then be reacted 'in methylene chloride tertiarybutyl alcohol solution with perchloric acid and N-bromoacetamide orN-iodosuccinimide to produce a reaction mixture from which 6'-fluoro-9a-bromo-l 1B,170Edil1YdfOXY-21-3C6tOXY- 1,4-pregnadiene-3,20-dione orthe corresponding 6-fluoro- 9u-i0d0 compound, respectively, can berecovered by precipitaltion with ice water and recrystallization fromacetone. The latter compounds can be reacted in acetone solutionwith'anhydrous potassium acetate at reflux temperature "to produce6-fluoro-9;3,1lfi-oxido-Uwhydroxy- 2l-acetoxy-l,4-pregnadiene-3,20-dionewhich is recoverable from the reaction mixture by chromatographicmethods' and can bepurified by recrystallization from a Skellysolve Bhexanehydrocarbons-acetone mixture. Reaction of the latter in methylenechloride solution with aqueous hydrogen fluoride at room temperature isproductive of 90,6-dlfil101'0-1113,17oL-dihYdIOXY 21acetoxy-1,4-pregnad'iene-3,20-dione. Substitution of aqueous hydrogenchloride, and carrying out the reaction at lower temperatures, e.g.,minus five degrees centigrade, is productive of 9a-chloro-6fluoro-11,8,17a-dihydroxy-2l-acetoxy- 1,4 pregnadiene-3,20-dione.

Substitution of 6-fluoro-115,17a-dihydroxy-21-acetoxy-4-pregnene-3,20-dione in the foregoing process is productive oftheanalogous resulting intermediate products, i.e., possessing the3-keto-'A structure in the A-ring of the steroidmolecule instead of the3-keto-A -structure, and results in 9a,6-difluoro-11,8,17a-dihydroxy-2lacetoxy-4- pregnene-3,20-dione and the corresponding 9oc-Chl01'0compound, respectively.

The compounds, 9a,6-dihalo-17u-hydroxy-21-acyloxy-4-pregnene-3,ll,20-trione (made in accordance with the process of theforegoing paragraphs, using as starting material6-halo-17a-hydroxy-21-acyloxy-4-pregnene-3,11, 20-trione) and9a,6-dihalo-11 8,17a-dihydroxy-21-acyloxy- 4-pregnene-3,20-dione, can beutilized as starting material for the corresponding9a,6-dihalo-17oz-hydroxy-21- acyloxy-l,4-pregnadiene-3,11,20-trione and9a,6-diha1o- 1113,17a-dihydroxy-2l-acyloxy 1,4 pregnadiene-3,20- dionebyfermentative l-dehydrogenation using Septomyxa or Fusarium organisms orby chemical l-dehydrogenation using selenium dioxide under theconditions previously described for the 1-dehydrogenation of CompoundVIII to Compound IX.

The 9a,6-difluoro-11,3,17a-dihydroxy 21 acetoxy-1,4-pregnadiene-3,20-dione or the 9a-chloro analogue thereof can be oxidizedwith chromic acid in accordance with known methods for convertingKendalls Compound F 21-esters to Kendalls Compound E 21-esters toproduce 9a,6-difluoro-17x-hydroxy 21 acetoxy-1,4-pregnadiene-3,11,20-trione and 9ot-ChlOI'O-6-fl1l0l0-l7oc-hYd1OXY-2l-3CB-toxyl,4-pregnadiene 3,11,20 trione, respectively. In a similar manner,the above compounds possessing the 3- keto-M-structu re in the A-ringcan be oxidized with chromic acid to produce9a,6-difluoro-17a-hydroxy-21- acetoxy-4-ipregnene-3J1,20 trione and9a-chloro 6-fluoro- 17a-hydroXy-2l-acetoxy-4+pregnene-3,1 1,20-trione,respectively.

V The foregoing dihalo compounds can be hydrolyzed to the corresponding21-hydroxy compounds according to known methods for hydrolyzing CompoundF Zl-acetate to Compound F, for example, in aqueous solution withpotassium bicarbonate under oxygen-free conditions. The free21-hydroxycompounds can be reesterified to produce the corresponding21-acyloxy compounds, using an ester-ifying agent, e.g., the anhydrideor acid halide of an organic carboxylic acid containing from one totwelve carbon atoms, inclusive, in the same manner as disclosed abovefor the monohalogenated analogs of hydrocortisone.

The foregoing Compounds VIILIX or X and their halo analogues, allcharacterized by the presence of a 6- fluoro or other 6-halo'substituen't, can exist in either the 60:- or the 6fi-epimeric'form.The foregoing process produces'a mixture in'whic'hthe 6/3-formpredominates. The 6a-epimer can be separated from the products bychromatographic or'fractional crystallization techniques known in theart. The compounds are useful, however, as such in the mixtures asproduced directly from the synthesis steps described. It is occasionallydesirable, nevertheless, to produce from the mixture'by separation or byepimerization the 6a-epimer which possesses even higheractivity than the6B-e'pimer. Conversion of the '6fi=epime'r or mixtures predominatingtherein can be accomplished by treatmentat temperatures 'of zero degreescenti-grade or slightly below in'an'essentially'anhydrous liquid mediumwith an anhydrous mineral acid, such as hydrochloric acid. The-mixtureshould be maintained at temperatures below zerodegrees centigr'ade, orat least below room temperature during the additionof the acid. Thereactionmixture can then be washed with successive-portions of'dilutealkali and water, and then dried and evaporated under reducedpressure.The 6u-fluoro, 6e-chloro or 6a-bromo products can be recovered from thecrude reaction product and purified by recrystallization.

The following examples are illustrative of the process and products ofthe present invention, but are not to be construed as limiting.

Example ].The 3-Ethylene Glycol Ketal of Methyl 3, 1 1 -Dike't0-5 ,6a-'Oxid0-'17(20)- [C is] Pregnen-21 -O:zte

To a solution of 5.0 grams of'the 3'-ethylene glycol ketal of methyl 3,1 1-diketo-4, 1 7 20 [cis] -pregnadien -2 l-oate, prepared in themanner described in US. Patent 2,707,184, in milliliters off-chloroformwas added a chilled solution of 1.9 grams of perbenzoic acid dissolvedin 31.5 milliliters'of chloroform. The solution was maintained ataboutfour degrees centigrade for 24 hours, and'then at room temperature'for72 hours. The solution was'then washed with a five percent aqueoussolution of sodium bicarbonate'and then with water. was separated, driedand the solvent distilled to give a residue of 5.3 grams of solid.Crystallization of this solid 'froin methanolgave 2.24 grams'ofproductrnelting at to degrees Centigrade and after two crystallizationsfrom methanol, there was obtained pure B-ethylene glycol ketal 'ofmethyl 3,11-diketo-5a,6a=oxido-17(20)- [cis]=pregnen-21'-oate melting at206 to 209 degrees centigrade "havingan [m1 of 37 degrees (CHCl andhaving the analysis given below: 7 V N g Calculated for C ,,H O C,69.20; H, 7.75. Found: C, 69.59;1-1, 7.81.

Example 2 Methyl 3,1 1 -Diket0-5u,6 3-Dihydr0xy 1 7 (20-All0pre'gnen-21-Oate and M ethyl 3,1 J-DiketO-Sa- Hydr0xy-6fi-F1u0r0-17 (20 -All0pregnene-21 -0ate To a solution of 1.73 grams of 3-ethyleneglycol ketal of methyl 3,11-diketo-5a,6a-oXido-17(20)-[cis]-pregnen-2l-oate in sixteen 'milliliters of methylene chloride was The chloroformlayer added six milliliters of 48 percent hydrofluoric acid. Theheterogenous mixture was stirerd for two hours, made slightly basic with300 milliliters of five percent sodium bicarbonate solution, andextracted with methylene chloride. The extract was washed, dried, andevaporated to dryness to give 1.62 grams of crude solid. Chromatographygave two fractions: (A) 481 milligrams eluted with methylene chlorideplus five percent acetone and (B) 921 milligrams eluted with methylenechloride plus ten and twenty percent acetone. Crystallization offraction (A) from acetone-Skeliysolve B hexanes gave 390 milligrams ofmethyl 3,11-diketo-5a-hydroxy-6fl-fiuoro- 17(20)-allopregnen-21-oate,melting point 254 to 260 degrees centigrade. The analytical samplemelted at 260 to 263 degrees Centigrade.

Analysis.Calculated for C H O F: F, 4.84. Found: F, 4.47.

Fraction (B) on crystallization from acetone-Skellysolve B hexanes gave470 milligrams of methyl 3,11-diketo-5a,6,B-dihydroxy-17(20)-allopregnen 21 oate, melting point 235 to 245degrees centigrade. The analytical sample melted at 245 to 248 degreescentigrade.

Aizalysis.Calculated for C H O C, 67.67; H, 7.74. Found: C, 67.91; H,7.62.

Following the procedure described in Example 2 but substituting hydrogenchloride at zero degrees centigrade for the hydrogen fluoride, there isthus produced methyl 3,11-diketo 50chydroxy-6B-chloro-17(20)-allopregnen- 21-oate, recoverable as a whitecrystalline solid from the reaction mixture by chromatography andcrystallization from acetone-Skellysolve B hexanes.

Example 3 .M ethyl 3,11-Diket-5ot-Hydroxy-fifl-Fluro-17(20)-All0pregnen-21-Oate 3-Eelllylene Ketal A mixture of 1.9 grams ofmethyl 3,11-diketo-a-hydroxy-65-fiuoro- 17(20)-allopregnen 21-oate, 59milligrams of p-toluene-sulfonic acid monohydrate and 31 milliliters ofdistilled ethylene glycol was added to 800 milliliters of benzene. Themixture was stirred and refluxed for two hours, with the condensatepassing through a Water trap to remove the water. After reflux themixture was cooled, washed with water and evaporated to dryness to givea crude solid which on recrystallization from acetone-Skellysolve Bhexanes gave 1.96 grams of methyl3,11-diketo-Sa-hydroxy-6B-fiuoro-17(20) allopregnen-Zl-oate 3-ethyleneketal, melting point 170 to 173 degrees Centigrade.

Following the above procedure, substituting other dihydric alcohols forethylene glycol, for example, 1,2-propylene glycol, 2,3-butanediol,1,3-butanediol and 2,3- pentanediol is productive of the respective3-alkylene ketals of methyl 3,11-diketo-5a-hydroxy-6fi-fluoro-17(20)-allopregnen-Zl-oate.

Example 4.5ez,11 3,21-Trihydr0xy-6fi-Flu0r0-17(20) A ll0pregnen-3-One 3Ethylene K etal To a solution of 1.96 grams of methyl3,11-diketo-5ahydroxy-6 3-fluoro-17(20)-allopregnen-21-oate 3-ethyleneketal in 850 milliliters of anhydrous ether was added 3.7 grams oflithium aluminum hydride and the mixture was stirred for a period of onehour. 200 milliliters of water was added slowly and the ether phaseseparated. The aqueous phase was extracted with ethyl acetate and theextracts added to the ether phase. The combined etherethyl acetatesolution was washed with water, dried and evaporated to dryness underreduced pressure. The crude solid residue was crystallized fromacetone-Skellysolve B hexanes to give 1.30 grams of5a,115,21-trihydroxy-6,B- fluoro-17(20)-allopregnen-3-one 3-ethyleneketal, melting point 197 to 205 degrees Centigrade. An additional 226milligrams was obtained from the mother liquor, melting point 175 to 185degrees Centigrade.

i 0 Example 5 .5 0a,] I,B-Dilzydr0xy-6fl-Flu0i'0-21-Acet0xy- 17(20)-All0pregnen-3-One 3-Ethylene Ketal The acetate was prepared by allowing0.87 gram of 501,115,21 trihydroxy 6,8 fluoro 17(20) allopregnen- 3-0ne3-ethylene ketal to stand overnight in ten milliliters of aceticanhydride and ten milliliters of pyridine. The solution was then pouredinto ice water to give 0.92 gram of 5a,1 1 fi-dihydroxy-dB-fluoro-Zl-acetoxy- 17 (20 allopregnen-3-one 3-ethylene ketal, melting point todegrees centigrade, which on recrystallization from acetone-SkellysolveB hexanes gave 0.77 gram, melting point 149 to 153 degrees centigrade.

Similarly, other 21-organic carboxylic esters of 50:,1113, 21trihydroxy-6B-fluoro-17(20)-allopregnen-3-one 3-ethylene ketals meprepared wherein the 21-acyloxy group is formyloxy, propionyloxy,butyryloxy, valeryloxy, hexanoyloxy, heptanoyloxy, octanoyloxy,benzoyloxy, phenylacetoxy, or the like, by contacting5a,11,8,21-trihydroxy- 6fl fiuoro-l7(20)-pregnen-3-one 3-ethylene ketalwith an appropriate acylating agent, e.g., the anhydride or acid halideof the selected acid in a solvent such as, for example, benzene,toluene, acetic acid, or the like. A convenient method of preparing the21-formyloxy ester consists in contacting 5a,1 1,8,21trihydroxy-6B-fluoro-17(20)-pregnen-3-one 3-ethylene ketal with formicacid in the presence of para-toluenesulfonic acid.

Example 6 .5 0a,] 1 5,1 7a-Trihydr0xy-6,3-Flu0r0-21 -Acetoxyallopregnane-3,20-Dione 3-Ethylene Ketal To a solution of 0.77gram of 5a,11/3-dihydroxy-6B- fluoro 21 acetoXy-l7 (20)-allopregnen-3-one 3-ethylene ketal in 35 milliliters of tertiary butylalcohol was added one milliliter of pyridine, 1.9 milliliters ofN-methyl morpholine oxide peroxide solution, and 13.1 milligrams ofosmium tetroxide (9.1 milliliters of tertiary butyl alcohol solutioncontaining 1.44 milligrams OsO per milliliter). The solution was stirredfor a period of 2.5 hours, fifteen milliliters of five percent sodiumhydrosulfite added, stirred for an additional ten minutes, 0.7 gram offinely ground synthetic magnesium silicate added, stirred for a periodof twenty minutes more and filtered. The filtrate was taken to drynessunder reduced pressure (below fifty degrees centrigrade) and the residuedissolved in methylene chloride, washed with water, dried and evaporatedto dryness. This residue was crystallized from acetone-Skellysolve Bhexanes to give 0.47 gram of 5 x,11,8,17atrihydroxy-Gfi-fiuoro-Zl-acetoxyallopregnane- 3,20-dione 3-ethyleneketal, melting point 220 to 228 degrees centrigrade.

Example 7.5 z,1 1[3,170t-Trihydroxy-6fi-Fluor0-21-Acetoxyallopregnane-3,20-Di0ne A solution of 0.47 gram of5a,11;3,17a-trihydroxy-6B- fluoro-21-acetoXyallopregnane-3,20-dione3-ethylene ketal in 35 milliliters of acetone and four milliliters of 1N sulfuric acid solution was gently boiled on the steam bath for tenminutes, cooled and neutralized with dilute sodium bicarbonate solution.Addition of water and cooling gave 0.33 gram of511,11,3,170c-1Iil1Yd10XY-6fi-fitl0l0-21-acetoxyallopregnane-3,20-dione, melting point 230 to 240 degreescentrigrade.

Example 8.6 8-Fluor0-1Jfi,17a-Dihydroxy-21 Acetoxy-4-Pregnene-3,20-Di0ne (6 8-Flu0r0hydr0c0rtis0ne Acetare) A solution of100 milligrams of 5u,11/3,17ct-trihydroxy-6B-fiuoro-21-acetoxyallopregnane-3,20-dione in 4.9 milliliters of aceticacid and 0.1 milliliter of water was refluxed for a period of one hour,cooled, diluted with fifty milliliters of water and evaporated todryness under reduced pressure. The residue was chromatographed overFlorisil (synthetic magnesium silicate) to give one fraction (77milligrams) eluted with methylene chloride plus ten 1 1. percentacetone. Crystallization from acetone-Skelly solve B hexanes gave 38milligrams of6,8-fiuro-11/8,17adihydroxy-21-acetoxy-4-pregnene-3,20-dione (6,8fluorohydrocortisone acetate), melting point 210 to 218 degreescentigrade. Infrared data and ultraviolet data are in agreement with thestructure.

Example 9.-65-Flu0r0-1 1 [3,1 7CL-DillYdlOXY-21ACE1OXZY-1,4-Pregnadiene-3,20-Dione A medium consisting of one percent dextrosehydrate, two percent cornsteep liquor of sixty percent solids andKalamazoo tap water was adjusted to pH 4.9 sodium hydroxide. The mediumwas steam sterilized at fifteen pounds pressure for thirty minutes,cooled, and then inoculated with a 24-hour growth, from spores, ofSeptomyxa aflinis, A.T.C.C. 6737. The medium was agitated, sparged withsterile air at the rate of one-tenth volume of air per volume of mediumper minute. At the end of 24 hours of fermentation at room temperature,the pH was about 7.4. To this culture there was added a solution of 6Bfiuoro 115,17u dihydroxy 21 acetoxy-4-pregnene- 3,20-dione(GB-fluorohydrocortisone acetate), dissolved in diethylformamide. Thesolution was prepared by dissolving five parts of the steroid in 100parts of the solid and adding about ten cc. of the solution per liter ofthe medium. Fermentation was continued for a period of 48 hourswhereupon the mycelium and beer were extracted thoroughly with methylenechloride. The extract was Washed with sodium bicarbonate solution andthen with water, dried and concentrated in vacuo to a slightly viscousresidue. The residue, after reacetylation with acetic anhydride inpyridine, was fractionated chromatographically and6B-fluoro-115,l'7a-dihydroxy-2l-acetoxy- 1,4-pregnadiene-3,20-dione wasrecovered as a lightcolored crystalline solid.

Following the procedure of Example 9, above, but substituting asstarting material9a,6-difluoro-11,8,17a-dihydroxy-21-acetoxy-4-pregnene-3,ZO-dione andreacetylating the recovered crude product, there was produced 9m,6-difluoro 11B,17a-dihydroxy-21-acetoxy-1,4-pregnadiene- 3,20-dione as awhite crystalline solid.

Example 10 Following the above procedure, but substituting the 6,8-fiuoro-11fl,17a dihydroxy 21 acetoxy-4-pregnane-3,20- dione(65-fluorohydrocortisone acetate) by6,8-luoro-l7ahydroxy-21-acetoxy-4-pregnene-3,11,20-trione(GB-fluorocortisone acetate) which was made by the oxidation of theproduct of Example 8 according to the methods disclosed in US. Patent2,751,402, yielded6fi-fluoro-17ahydroxy-2l-acetoxy-1,4-pregnadiene-3,l1,20-trione as acrystalline product.

In accordance with the foregoing examples, methyl 3, 1 1-diketo-5a-hydroxy-6 ,G-chloro-17 (20) -allopre gnen-2 1- oate can be substitutedin the process of Example 3 to produce the 3-ketal thereof. The lattercompound can then be subjected to reduction according to the process ofExample 4 to produce 5a,115,2l-trihydroxy-Gfi-chloro- 17(20)-allopregnen-3-one 3-ethylene ketal and the latter compoundconverted to its 2l-acetate or other organic acid ester by the procedureof Example 5. subjecting the thusproduced511,11B-dihydroxy-fl-chloro-Z1-acetoxy-17(2O)- allopregnen-B-one3-ethylene ketal to oxidative hydroxylation in accordance with Example6, followed by hydrolysis of the 3-ketal group by the procedure ofExample 7, and further dehydrating in accordance with the procedure ofExample 8, is productive of 6,8-chloro-1lfl,17a-dihydroxy-21-acetoxy-4-pregnene-3,20-dione (6,6 chlorohydrocortisone acetate). Thelatter compound, when substituted in the procedure of Example 9, isproductive of 6,8-chloro- 115,170; dihydroxy 21 acetoxy1,4-pregnadiene-3,20- dione. Furthermore, 6fi-chlorohydrocortisoneacetate can be oxidized to 6B-cl1lorocortisone acetate as shown abovefor the 6/3-fluoro analogue. Substitution of 6,8- chlorocortisoneacetate in the process of Example 9 is 12 productive of 6,8-chloro-118,17a-dihydroxy-21-acetoxy- 1,4-pregnadiene-3,20-dione.

As previously described, the 21-acyloxy products of this invention,e.g., the products of Examples 9 and 10 or the corresponding 6 8-chloroanalogues thereof, can be hydrolyzed to the corresponding free21-alcohols by the hydrolysis procedure described above. In thehydrolysis procedure, as previously stated, it is desirous to employ anoxygen-free solution and oxygen-free conditions and to use a molarexcess of an alkali-metal bicarbonate such as potassium bicarbonate as ahydrolytic agent. The temperature is preferably held lower, i.e.,between ten and about thirty degrees centigrade, and the hydrolyzedsolution subsequently neutralized with an acid such as acetic acid.

The compounds disclosed in Examples 9 and 10 and their 6B-chloroanalogues possess valuable anti-rheumatoid arthritic, anti-inflammatoryactivity and can be used and administered as disclosed in the foregoingdescription.

Example J1.Is0merizati0n of 65- to 6oc-Flu0r0hydr0- cortisone 21-AcetateA solution of 0.132 gram of 65-fluorohydrocortisone 2l-acetate in twelvemilliliters of chloroform and 0.1 milliliter of absolute alcohol wascooled to minus ten degrees centigrade in an ice-salt bath and a streamof anhydrous hydrochloric acid was gently bubbled through the solutionfor 2.5 hours while the temperature was maintained between minus fiveand minus fifteen degrees centigrade. The solution was then diluted with25 milli liters of chloroform, washed with dilute sodium bicarbonate andwater, dried over anhydrous sodium sulfate, and evaporated to drynessunder reduced pressure at sixty degrees centigrade or less.Crystallization of the residue from acetone-Skellysolve B gave 42milligrams of product, 6a-fiuorohydrocortisone 21-acetate, melting point203 to 210 degrees centigrade.

In a similar manner, 6B-fluoro-11,8,17a-dihydroxy-21-acetoxy-l,4-pregnadiene-3,ZO-dione is converted to 60- fluoro 11,3,17udihydroxy-21-acetoxy--1,4-pregnadiene- 3,20-dione, and65,9u-difluoro-11[3,17a-dihydroxy-21-acetoxy-l,4-pregnadiene-3,20-dioneis converted to 6oc,9oL-d1 fluoro 11,8,l7adihydroxy-21-acetoxy-1,4-pregnadiene- 3,20-dione.

It is to be understood that the invention is not to be limited to theexact details of operation or exact compounds shown and described, asobvious modifications and equivalents will be apparent to one skilled inthe art, and the invention is therefore to be limited only by the scopeof the appended claims.

I claim:

1. 6 chloro 11 oxygenated-l7ot,2l-dihydroxy-1,4- pregnadiene-3,20-dioneof the following formula:

wherein R is a member selected from the group consisting of fl-hydroxyand keto; and the 2l-acylates thereof wherein the acyl radical is thatof a hydrocarbon carboxylic acid containing from one to twelve carbonatoms, inclusive.

2. 6-chloro-l-dehydrohydrocortisone.

3. 6-chloro-1-dehydrocortisone.

OOQQUI CH OH 14 9. A compound represented by the formula:

(EHZOH 0:0 5 I OH OH] 15 References Cited in the file of this patentUNITED STATES PATENTS 2,837,464 Nobile June 3, 1958 2,881,168 Spero Apr.7, 1959

1. 6 - CHLORO - 11 -OXYGENATED-17A,21-DIHYDROXY-1,4PREGNADIENE-3,20-DIONE OF THE FOLLOWINGFORMULA: